1. Field of the Invention
The present invention relates to a method for preparing a natural pharmaceutical composition that contains both statin acting as a cholesterol biosynthesis inhibitor, and a CoQ10 compound as a substance that inhibits factors causing complications involved in oral administration of the statin, thus exhibiting multi-physiological active functions such as anti-arteriosclerosis and cardiac function reinforcement, through fermentation using Monascus sp. and natural substances having pharmaceutical efficacies.
2. Description of the Related Art
Coenzyme Q10 (ubiquinone, CoQ10) is a quinone substance similar to fat-soluble vitamins, which are present throughout human tissues. The major biochemical function of Coenzyme Q10 is as an electron carrier in intracellular energy (ATP) metabolism in cellular respiration of mitochondriona (Sarah L. Molyneux et al., Journal of the American College of Cardiology, 52, 1435-1441, 2008). Also, Coenzyme Q10 functions as an endogenous antioxidant preventing oxidative damage of cellular membranes and removing reactive oxygen species and the like. CoQ10 is vigorously biosynthesized in the twenties in humans and biosynthesis thereof decreases with age. Accordingly, research into industrial mass-production of CoQ10 and utilization of the same in a variety of applications including medicines, functional health functional foods, and cosmetics has been conducted (Littarru et al., Nutr. Res., 42, 291-305, 1972; Khatta M et al., Ann Intern Med, 132, 636-640, 2000). 90% or more of CoQ10 present in human serum and organism tissues is a reduced form of ubiquinol (ubiquinol, CoQ10H2) which is a potent fat-soluble antioxidant. A ratio of CoQ10H2:CoQ10 and a decrease in serum CoQ10 are related to oxidative stress. Oxidative stress defined by disequilibrium between prooxidants and antioxidants is known to be a major risk factor related to pathologic conditions such as cardiovascular disease, diabetes and cancer as well as aging (S. Yamashita et al., Anal Biochem, 250, 66, 1997; D. Steinberg et al., Engld J Med, 320, 915, 1989). In particular, a ratio of ubiquinol and ubiquinone, both of which are CoQ10 is commonly used as a reference value in a great deal of research associated with the prevention, diagnosis and treatment of oxidative damage. Oxidation of plasma lipoprotein is a major cause of arteriosclerosis as well as other diseases associated with free radicals such as Parkinson's disease, Alzheimer's disease or beta-thalacemia. In this case, ubiquinol removes peroxy radicals, reduces alpha-tocopheryl radicals and thereby prevents oxidation of LDL. In fact, as compared to alpha-tocopherols acting as plasma antioxidants, ubiquinol is an endogenous antioxidant that first reacts with plasma, regardless of concentration, when the plasma is exposed to the antioxidant. Accordingly, use of supplements in order to reinforce CoQ10 is beneficial in the enhancement of human health. Intake of CoQ10 via foods or food supplements increases a level of CoQ10 in blood and helps reinforce antioxidant functions, thus preventing onset of major diseases caused by cellular oxidative damage.
A great deal of research to identify relation between CoQ10 deficiency, disease conditions, clinical improvement after administration of CoQ10 and the like has been made. First of all, it was demonstrated that CoQ10 is efficacious in lengthening statin treatment without side effects and maintaining an essential cellular bio-energy amount when it is supplemented to hypercholesterinemia patients who should take statin drugs for a long period of time. Statin that was first developed in 1987 has contributed to decrease cardiovascular diseases, as a first-generation cholesterol inhibitor. However, commonly, humans expect to develop a second-generation cholesterol suppressor that has more specific actions while not interfering with biosynthesis of other compounds, like CoQ10. Accordingly, if drugs (supplements) consisting of natural substances containing predetermined concentrations or higher of both statin and CoQ10 which are natural anti-cholesterol components have been developed, side effects of statin which is an anti-cholesterol drug are removed through additional or synergistic treatment effects between these compounds and health benefits can be obtained through bioactivity enhancement of CoQ10.
All over the world, coronary artery diseases are considered to be major mortality factors and, in particular, hypercholesterinemia is considered to be the most risk factor.
⅔ or higher of total cholesterol in humans is obtained by biosynthesis. In the biosynthesis, a step on which an HMG-CoA reductase (3-hydroxy-methyl-3-glutaryl-coenzyme reductase) acts is an initial step and, at the same time, is a step that limits a speed of synthesis route (FIG. 3).
HMG-CoA reductase inhibitors globally used in recent years, that is, statin (or mevinolin, lovastatin, monacolin K, Mevacor, C24H36O5) drugs have been used for a long period of time in order to effectively treat hypercholesterinemia and prevent death caused by cardiovascular diseases. Statin drugs are considerably potent substances that directly inhibit biosynthesis of endogenous cholesterol corresponding to 80% of total cholesterol, but statin drugs are classified into special medicines that are sold in the presence of a medical prescription, since an activation process in which the ring of molecular structures is opened by hydrolase (carboxyesterase) causes damage to the liver and kidneys, as well as side effects such as myopathy (Thomson P D et al., JAMA. 289: 1681-1690 2003). However, statins inhibit HMG-CoA reductases and thereby reduce production of cholesterol, while they have the same biosynthesis route as ubiquinone, which are in-vivo electron and quantum carriers and thus affect biosynthesis of cholesterol as well as CoQ10, resulting in deterioration in yields of two substances (FIG. 3). That is, unfortunately, potential impacts of statins at a mevalonate-level are unspecific and, as a result, statins inhibit biosynthesis of various sterol isoprenoids including CoQ10 and dolichol. Accordingly, patients who take statin for a long period of time are known to suffer from side effects such as myalgia, since a level of biosynthesized CoQ10, which plays a role as an essential factor in the energy production of mitochondriona, in the blood is increased. That is, use of statins decreases a concentration of CoQ10 in plasma, increases a ratio of lactic acid to pyruvic acid and, furthermore, deteriorates functions of mitochondriona respiration system. As a consequence, deficiency of energy production increases a fatigue degree of muscles and thus decreases aerobic capacity (respiration capacity).
Accordingly, use of statin drugs for a long period of time is found to deteriorate cardiac function and blood circulation due to decrease in level of CoQ10 in the blood, thus resulting in serious side effects. That is, statin drugs are reported to cause side effects of digestive systems (such as constipation, diarrhea, dyspepsia, abdominal inflation and abdominal pain) and side effects of nervous systems (such as dizziness and headache) and side effects such as skin rashes and visual disturbances. In particular, side effects such as liver failures are proved to be serious to an extent that patients stop taking their statin drugs, and development of a method for maintaining a concentration of CoQ10 in the blood of statin users at a predetermined level was demanded. In this regard, U.S. Pat. No. 5,082,650 discloses that a correlation between variation in concentration of CoQ10 in the blood and side effects in various cases is observed with respect to patients, to which statin drugs (MEVACOR) are applied, a correlation between the concentration of CoQ10 in the blood, and reduction and removal of side effects through supplementing with CoQ10 is discovered and prescription of CoQ10 is also required for users of statin drugs. Accordingly, it is found that statin drugs used in order to maintain the concentration of cholesterol in the blood should be administered in combination with supplement of CoQ10 (200 mg per day) to prevent serious side effects generated in the heart and liver.
For this purpose, similar research results to execute the US patent are disclosed in Korean Patent Laid-open No. 2009-28983. This patent discloses a multi-therapeutic pharmaceutical composition for oral administration, containing both statin drugs and coenzyme 10 as synthetic medicines in order to prevent side effects of the statin drugs which are synthetic medicines for treating hyperlipidemia, wherein the composition contains a great amount of a solubilizer such as polyethylene glycol to increase solubility of water-insoluble lactone-type synthetic statin, a surfactant, an oil to provide an emulsion, a sodium or calcium antioxidant to secure stability, a pH adjuster or the like.
However, statin drugs and CoQ10, main components of the substance, are artificially synthesized compounds, which are water-insoluble and thus have problems of solubility in the intestine. In particular, calcium salt medicines, pH, of which is adjusted to 6.7, have a problem of considerably low solubility.
However, the present invention is designed under the assumption that all the problems can be solved, if an acid-form natural statin compound having a pH of 6.5 to 7.0 is bio-synthesized by natural fermentation.
To date, Monascus sp. (red yeast rice red koji) is prepared by inoculating rice with the genus Monascus (red mold fungi), which has been used as natural food colorings or products or substances for digestion promotion and blood stream improvement in various East Asian nations including China for a long time. In particular, it was reported that natural statins (HMG-CoA reductase inhibitors) which are secondary metabolites produced by Monascus sp. potently inhibit HMG-CoA reductases, cholesterol biosynthesis enzymes, and thus exhibit functionalities of various bioactivities, based on deterioration in blood-lipid concentration and cholesterol biosynthesis (Wang I K et al., J. Agri. Food Chem. 48: 3183-3189 2000; Endo A. et al., J. Antibiotechnol. 38: 420-422 1985; Manzoni M & Rollini M. App. Microbiol. Biotechnol. 58: 555-564 2002; Wei W. et al., J. Nutri. Biochem. 14: 314-318 2003). Since then, Monascus sp. has been widely used as an herbal medicine in Chinese “medicine”.
Meanwhile, to date, submerged fermentation of liquid culture has generally been used for production of Monascus sp. cultures. Optimum fermentation conditions to obtain a pigment component and statin (mevinolin) as functional metabolites, strain search, and preparation of functional products containing Monascus sp. have been studied. Also, solid fermentation is also used in order to obtain functional components including pigments, but it is known that a main cereal of used medium is rice, barley and wheat are also used, and bean (white bean) is unsuitable for use in a Monascus sp. solid medium due to high protein content. However, the inventors of the present invention established fermentation conditions using bean, as a substrate for a solid medium, instead of rice, developed a method for producing statins derived from Monascus sp. fermented with bean, and obtained first patents in Korea as well as other nations (Korean Patent No. 0734612). However, this invention only aims at successively producing Monascus sp. fermented with bean without producing citrinin that acts as a kidney or liver toxin by fermenting Monascus sp., using bean, instead of rice, for preparation of a solid medium for statin (mevinolin) biosynthesis.
In order to overcome the limitations of medium preparation, the present inventors provide a safe and improved natural pharmaceutical composition for preventing and treating hyperlipemia wherein a biosynthesized statin amount is increased by 10% or more and a great amount of CoQ10 is simultaneously accumulated, by remarkably changing a composition of a seed culture. Furthermore, the present inventors attempt to identify biosynthesis of Monascus sp.-derived citrinin that acts as a toxic component that causes functional disorders of kidney and liver, as mold metabolites in in vivo pharmaceutical functional mechanisms.
Conventional methods associated with the specific medium composition of Monascus sp. of the present invention and simultaneous biosynthesis of the multi-physiologically active substance of the present invention include Korean Patent Laid-open No. 2005-13188 that discloses a composition containing statin for treating leukemia, Korean Patent Laid-open No. 2007-7022964 that discloses treatment using statin, omega-3 fatty acid and a combination thereof, Korean Patent Laid-open No. 2007-7003811 that discloses a novel statin pharmaceutical composition and a method for treating diseases associated therewith, Korean Patent Laid-open No. 2007-7000831 that discloses a composition containing statin and a bronchodilator, Korean Patent Laid-open No. 2006-7015689 that discloses a composition for treating hypocholesterolemia containing a statin and a de-gassing agent, and Korean Patent Laid-open No. 2009-7004090 that discloses a pharmaceutical composition containing statin-capsulated nanoparticles. However, it could not identified that the conventional patents deny novelty and inventive step of the present invention, with respect to liquid culture media associated with simultaneous biosynthesis of statin and CoQ10, as natural combined physiologically active substances, and simultaneous production methods for these substances.